equilibrium concentration of the weak acid and

O U T L I N E its conjugate base.

I. Introduction C. Both ● Ratio of concentrations of weak acid and
Reagents conjugate can be calculated
II. Preparation of are in ● Respective masses can then be
Buffers in the Aqueous determined
Laboratory Form
A. Weak D. Only One A. Weak Acids and their Conjugate Bases are
Acids and Reagent Solids
their is
Conjugate Available Step 1
Bases are Obtain necessary details such as target pH,
Solids volume, and concentration of the buffer to be
B. One of the prepared.
Reagents
is in Step 2
Aqueous Choose the best weak acid to be used as a buffer:
Form ● Weak acid with nearest pKa value to
target pH
● Buffers work properly when pH of the
I. INTRODUCTION
buffer solution is within the buffering
Buffers Solutions that resist drastic changes range: pKa + 1 of the weak acid
in pH levels when strong acid/base ● HHE also works best around this region
is added → significant amounts of weak acid and
conjugate base are present
● Composed of weak acid/base and its
conjugate base/acid
Table 1. pKa Values of Some Common Weak
○ Counteracts the H+ or OH- given
Acids used as Buffers.
off by the addition of small
amounts of acids and bases Weak
pK1 pK2 pK3
● Physiologically important because they Acid
maintain the pH necessary for living
organisms to thrive Acetic
4.74 - -
Acid
II. PREPARATION OF BUFFERS IN THE
Carbonic
LABORATORY 6.10 10.4 -
Acid

Henderson-Hasselbach Equation Phosphor

● pH = -log[H+] 1.97 7.00 12.5
ic Acid
● pKa = -log(Ka), where Ka = acid
equilibrium constant Citric
3.10 4.10 6.40
● [HA]: M of weak acid; [A-]: M of conjugate Acid
base
Example: In the preparation of a 500 mL of 0.1 M
buffer with pH 7.4, which weak acid should be
used?
Equation 1. Henderson-Hasselbalch Equation. Answer: Phosphoric acid with pK2 of 7.0 which is
near the target pH of 7.4
pH of a solution can be obtained by looking up
the Ka value of the weak acid involved, and the

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Step 3 Buffering Capacity: dependent on the
Use HHE to calculate the ratio between the concentration of the buffer components
conjugate base and the weak acid. ● Buffer containing greater amounts of
weak acid and conjugate base → longer
Example: Second acid dissociation constant (pK2) buffering action → higher buffering
will be used. Following the dissociation pattern capacity
for phosphoric acid (H3PO4), chemical species
present at the target pH will be H2PO4- and Step 5
HPO42-. Calculate the masses of the reagents as they are
in solid form.

Phosphate species are usually sold as sodium

salts NaH2PO4 (sodium dihydrogen phosphate,
MW = 119.98 g/mol) and Na2HPO4 (sodium
hydrogen phosphate, MW = 141.96 g/mol).
Substituting the target pH of 7.4, pK2 of 7.0, and
● Target volume (500 mL = 0.5 L) will be
the chemical species in the HHE, the ratio of
needed to calculate the masses.
conjugate base to its weak acid can be solved:

Step 6
Weigh the reagents and mix them in a beaker.
Add distilled water until all solid particles are
dissolved. Volume added must not be 500 mL
right away.
For every mol of H2PO4-, there is 2.51 times more
HPO42- → [H2PO4- ] must be less than [HPO42-] Step 7
because target pH value of 7.4 was over pK2 After the dissolution, measure pH using the pH
value of 7.0 (more basic than original since meter.
conjugate base is higher in concentration) ● If higher than target pH, add 0.1 M HCl
dropwise until desired pH.
Step 4 ● If lower than target pH, add 0.1 M NaOH
Using the target buffer concentration (0.1 M), the dropwise until desired pH.
following equation can be established. Only This discrepancy in pH is a result of the
H2PO4- and HPO42- are present at target pH → dependency of Ka on temperature, where Ka
[H3PO4] and [PO43-] are essentially negligible values are usually given at 25oC.

Step 8
Transfer the solution with adjusted pH to
appropriate volumetric flask then fill the
distilled water until the mark. Mix thoroughly
before use or store at 4oC until use.
[HPO42-] can be calculated after getting [H2PO4-].
For confirmation, the ratio of the conjugate base
to the weak acid should be 2.51.

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 B. One of the Reagents is in Aqueous Form where M is the proton (H+) concentration (in
Steps are almost the same with A except for molarity, M) and V is the volume (L or mL); [H+] =
Step 5 10-pH
● This preparation usually happens when
weak acids are only available in aqueous Target volume: 250 mL → summation of the
form e.g., phosphoric acid (H3PO4), acetic volumes of the buffer solutions of pH 6.5 and 7.5
acid (CH3COOH), etc. or when using
already prepared solutions.
● Volume of the reagent (given known
concentration of stock solution) is
computed instead of mass. Substituting the equation and other relevant data
to the initial equation:
Example: For a particular buffer (500 mL), you
calculated its components to have the following
concentrations: [H3PO4] = 0.025 M, [H2PO4-] =
0.075 M. How do we make the buffer when the
stock reagents available are: 5.0 M H3PO4 (liquid)
and NaH2PO4 salt (FW= 119.98 g/mol). To prepare the 250.0 mL of a 0.10 M phosphate
buffer at pH 7.0, 60.1 mL of 0.10 M phosphate
buffer (pH 6.5) and 189.9 mL of 0.10 M
phosphate buffer (pH 7.5) must be mixed.
● 0.10 M concentrations from pH 6.5 and
7.6 buffer solutions will not matter
because the same number of phosphate
species will be obtained after dilution →
final buffer solution will still have 0.10 M
Hence, 4.50 g of NaH2PO4 and 2.5 mL of the 5.0 of phosphate species as its concentration
M H3PO4 will be mixed and the rest of the
process (Steps 6 to 8) will follow. D. Only One Reagent is Available
Most time-consuming of the preparations
C. Both Reagents are in Aqueous Form
Volumes will be measured and mixed in this type Example: You are tasked to prepare 500 mL of
of buffer preparation. 0.1 M buffer with pH 7.4. Unfortunately, only the
● Calculation will use a modified dilution NaH2PO4 (MW = 119.98 g/mol) is present in the
equation laboratory. How will you prepare the buffer from
● Usually done when buffers of the same scratch?
concentration but different pH are already
prepared Calculate the mass directly from NaH2PO4

Example: You were tasked to prepare a 0.10 M

phosphate buffer at pH 7.0. Unfortunately, only
0.10 M phosphate buffer solutions of pH 6.5 and
7.5 are available. How much of each buffer
solution must be mixed to prepare 250.0 ml of This means that 6.00 g of NaH2PO4 is needed to
the phosphate buffer at pH 7.0? produce 0.1 M of the needed phosphate
species. When dissolved, the pH will still be very
far from the target pH as compared to when both
weak acid and its conjugate are present.

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Conjugate bases can be derived from weak acids,
as in the case of H2PO4 where pK2 = 7.4:

From the HHE, we compute that we need 0.05 M

of H2PO4- and 0.05 M of HPO42- in the final buffer
to get a pH of 7.4. How do we get HPO42-
species from H2PO4-?

● NaOH + H2PO4- → HPO42-

● 1 mol of the conjugate base is created for
every 1 mol of weak acid and NaOH
● Thus, 0.05 M of NaOH + H2PO4- solution
→ 0.05 M HPO42- (NaOH reacts with
equimolar of H2PO4-)
○ Since initial concentration of
H2PO4- is 0.1 M, 0.05 M of H2PO4-
(weak acid) remains after reaction
with 0.05 M NaOH

In the resulting buffer solution, we have 0.05 M

each of the conjugate base, HPO42-, and the weak
acid, H2PO4- → buffer is at desired pH and
concentration

pH adjustment of the solution is time-consuming

as buffers resist drastic changes in pH

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